TY - JOUR
T1 - The effect of elevating extracellular CaCl2
T2 - Important considerations for tissue engineering applications
AU - Jaworska, Kayley
AU - Senior, Jessica
AU - Bruning-Richardson, Anke
AU - Smith, Alan
N1 - Funding Information:
The authors would like to thank the EPSRC Doctoral Training Partnership (EP/T51813X/1) and BBSRC 3DBioNet (MR/R025762/1) for funding and supporting this work.
Publisher Copyright:
© 2024 The Authors
PY - 2024/12/1
Y1 - 2024/12/1
N2 - Polysaccharides such as sodium alginate, pectin and gellan gum are widely used biomaterials, for their ability to easily form hydrogels in the presence of divalent metal ions, such as calcium - a process often cited as a mild crosslinking mechanism. However, when using these materials as substrates for tissue engineering, there is a lack of extensive studies that investigate the impact of elevated calcium concentrations on cell health and behaviour. In this study, we performed an in-depth exploration to understand the potential effects of raising extracellular CaCl
2 on cell viability, proliferation, morphology and migration. We used an established glioblastoma (GBM) cell line (U251), human dermal fibroblasts (HDF), and murine osteoblasts (MC3T3) to assess the consequences of using CaCl
2 in tissue engineered models to help reevaluate biomaterial suitability and enhance standardisation practices in the field of tissue engineering. Our findings revealed that the addition of CaCl
2 induced notable morphological changes in GBM cells when cultured in 3D hydrogels with excess CaCl
2 added, leading to a transition from mesenchymal to amoeboid phenotypes, even at a concentration as low as 8 mM. Furthermore, cell viability was reduced in a concentration-dependent manner across all cell types, and migration was also affected. Despite the widespread use of high CaCl
2 concentrations to facilitate scaffold gelation, our research unveils that there can be significant risks to cell viability, proliferation, morphology, and migration when such practices are not preceded by cell line-specific experimentation and thorough standardization procedures. This highlights the importance of careful consideration and optimisation of CaCl
2 concentration when used as a crosslinking agent for hydrogels intended for use in tissue engineering applications that demand accurate recapitulation of cellular responses and physiological conditions.
AB - Polysaccharides such as sodium alginate, pectin and gellan gum are widely used biomaterials, for their ability to easily form hydrogels in the presence of divalent metal ions, such as calcium - a process often cited as a mild crosslinking mechanism. However, when using these materials as substrates for tissue engineering, there is a lack of extensive studies that investigate the impact of elevated calcium concentrations on cell health and behaviour. In this study, we performed an in-depth exploration to understand the potential effects of raising extracellular CaCl
2 on cell viability, proliferation, morphology and migration. We used an established glioblastoma (GBM) cell line (U251), human dermal fibroblasts (HDF), and murine osteoblasts (MC3T3) to assess the consequences of using CaCl
2 in tissue engineered models to help reevaluate biomaterial suitability and enhance standardisation practices in the field of tissue engineering. Our findings revealed that the addition of CaCl
2 induced notable morphological changes in GBM cells when cultured in 3D hydrogels with excess CaCl
2 added, leading to a transition from mesenchymal to amoeboid phenotypes, even at a concentration as low as 8 mM. Furthermore, cell viability was reduced in a concentration-dependent manner across all cell types, and migration was also affected. Despite the widespread use of high CaCl
2 concentrations to facilitate scaffold gelation, our research unveils that there can be significant risks to cell viability, proliferation, morphology, and migration when such practices are not preceded by cell line-specific experimentation and thorough standardization procedures. This highlights the importance of careful consideration and optimisation of CaCl
2 concentration when used as a crosslinking agent for hydrogels intended for use in tissue engineering applications that demand accurate recapitulation of cellular responses and physiological conditions.
KW - calcium biocompatibility
KW - hydrogels
KW - crosslinking
KW - tissue engineering
KW - glioblastoma
KW - biomaterials
KW - Calcium
KW - Hydrogels
KW - Tissue engineering
KW - Glioblastoma
KW - Crosslinking
KW - Biomaterials
KW - Biocompatibility
UR - http://www.scopus.com/inward/record.url?scp=85209664063&partnerID=8YFLogxK
U2 - 10.1016/j.tice.2024.102615
DO - 10.1016/j.tice.2024.102615
M3 - Article
VL - 91
JO - Tissue and Cell
JF - Tissue and Cell
SN - 0040-8166
M1 - 102615
ER -
